Immortalized Retinal Müller Cell Line rMC-1
INVENTION: Müller cells are the most abundant non-neuronal cells in the vertebrate retina. It is known that they perform a variety of functions that support the activities of retinal neurons, but there has been little progress in elucidating the mechanisms underlying these functions. Primary cultures have provided useful insight into Müller cell biology, biochemistry, development, and electrophysiology, but their utility is somewhat limited because the culture size is quite small unless a very large number of retinas are used. Furthermore, problems are frequently encountered with senescence and with contamination by astrocytes and microglia. An immortalized cell line expressing the Müller cell phenotype has now been established by transformation with SV40. Immunochemical studies have shown that this new cell line (rMC-1) expresses (GFA)P, which is a marker for reactive gliosis, and CRALBP, a marker for Müller cells in the adult retina. In transient transfection assays, promoter proximal sequences of the CRALBP gene stimulated expression of a luciferase reporter gene. The rMC-1 cell line will significantly enhance our understanding of the interaction of Müller cells with endothelial cells and neurons. The availability of an immortalized cell line should also facilitate studies of gene regulation by circumventing the problems of low cell number and low transfection efficiency encounter with primary cell cultures.
FIELD OF APPLICATION: Research in mammalian retinal function, particularly in the areas of regulation of gene expression and cell-cell interactions.
ADVANTAGES: Müller cells must be isolated from a large number of retinas in order to obtain a sufficiently large cell population. This is time-consuming and costly, and resulting primary cultures are small, have a limited life span, and are often contaminated with astrocytes and microglia. An immortalized cell line with the Müller cell phenotype thus provides a valuable research tool for studies of the mammalian retina.
Vijay P. Sarthy
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